Simultaneous optical imaging of intracellular Cl− in neurons in different layers of rat neocortical slices: advantages and limitations

Two-photon chloride imaging in neurons of brain slices

“…3A). The washin of the calibration solution containing 10 M of nigericin and tributyltin chloride caused an immediate reduction of the MQAE-mediated fluorescence by 19€7.8% (n=42 cells), probably because of photon absorption caused by the drugs [16]. This reduction precluded the calibration of resting intracellular Cllevels.…”

“…The major disadvantage of all quinoline-based indicator dyes is, however, that they are prone to unusually strong bleaching [24,39]. This restricts the duration of the measurements and allows only a low data acquisition rate (0.2-2 frames per minute) [16,24,41].…”

The Chloride Transporter Na+-K+-Cl−Cotransporter Isoform-1 Contributes to Intracellular Chloride Increases afterIn VitroIschemia

“…Finally, Clomeleon absorbs and emits light at visible wavelengths, allowing Clomeleon to be used in conjunction with the Cl Ϫ transport inhibitors bumetanide and furosemide, which absorb UV light and quench MEQ (Fukuda et al, 1998 ] i in adult rat hippocampal slices subjected to OGD rise in [Cl Ϫ ] i . It is likely that OGD activates many pathways for Cl Ϫ entry.…”

Kinetic Properties of Cl−Uptake Mediated by Na+-Dependent K+-2Cl−Cotransport in Immature Rat Neocortical Neurons

“…3C), a [Cl Ϫ ] i of 29.2 Ϯ 1.4 mM was calculated using the Stern-Volmer equation. Four slices were preincubated with 100 M bumetanide for 1-3.5 h, because the absorption and emission spectra of bumetanide interfere directly with MEQ fluorescence (Fukuda et al, 1998). Subsequent experiments in bumetanide-free solutions revealed that in these slices, the ratio F 0 /F rest was reduced to 1.52 Ϯ 0.03 (n ϭ 36 cells from four slices) (Fig.…”

“…Slices were loaded with the Cl Ϫ -sensitive fluorophore MEQ by incubating them for 60 min in 370 M 6-methoxy-N-ethyl-1,2-dihydroquinoline (diH-MEQ). Preparation of diH-MEQ and loading procedure were described previously in detail (Fukuda et al, 1998). The slices were simultaneously loaded with the volume marker calcein (10 g/ml in the presence of 0.01% pluronic F127).…”